Separation and formulation of bioactive fraction and subfraction from camel urine work as anticancer agent

ABSTRACT

A pharmaceutical composition includes an effective amount of bio-active fraction PMF or subfraction PMFK obtained from the lyophilized camel urine or cow urine which coded in publications with PM701, that is used as anti-cancer drug selected from an anticancer compound PM701, which are targeted selectively the cancer cells without affecting the normal cells.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to an absolutely novel use of camel urinefor medicinal purposes and also relates to the preparation of it bylyophilization as a drug that called PM701 or fractionating it to activefractions that are coded as PMF and PMFK. More specifically, the presentinvention relates to isolating the bio-active fraction PMF and the mosteffective subfraction PMFK from the lyophilized PM701 of the adultsingle-humped Arabian camel, species camelus dromedarius and theabsolutely novel use of these bio-active fractions as a selectiveanti-cancer agent. The fractionation will be implicated in reducing thedosage of the whole lyophilized urine and increasing its efficacy. Theinvention also relates to the preparation of a novel pharmaceuticalcomposition (capsules) comprising an effective amount of the bio-activefraction PMF or the bioactive subfraction in a composition of 435mg/capsule with a range 7 or 5 capsules/day respectively, which aretargeted to the human cancer tissues through their anti-proliferativeand apoptotic activities without harming the other normal tissues.

2. Description of the Related Art

Cancers are uncontrolled cell proliferations that result from theaccumulation of genetic changes in cells endowed with proliferativepotential. After a variable latency period during which they areclinically silent, the malignant cells progress to aggressive invasiveand metastatic stages with tumor formation and wide-spread disseminationthroughout the body.

Despite important advances in treatment, cancers still account for 28%of death in Western countries, and more than this ratio in othercountries. Treatment of cancer has relied mainly on surgery,chemotherapy, radiotherapy and more recently immunotherapy. However, forthe most frequent types of cancers (lung, breast, colo-rectal and theleukemias) complete remission and cure has not been achieved. Therefore,the development of new approaches for treating cancer patients is highlydesirable and critically needed particularly for those patients whosedisease has progressed to a metastatic stage and are refractory tostandard chemotherapy.

The world health organization published that cancer is a leading causeof death worldwide. From a total of 58 million deaths worldwide in 2005,cancer accounts for 7.6 million (or 13%) of all deaths, in 2005 cancerkilled approximately 12,000 people in Saudi Arabia, as shown in FIG. 1,with 8,000 of those people were under the age of 70.

In Prophet Mohamed Medicine (peace upon him), camel urine is suggestedfor drinking to improve some symptoms mainly associated with tumorformation in the body. Therefore, camel urine has been used in theIslamic and Arab world, sold and distributed in different size bottles.But it is never scientifically tested. The applicant considered it isworthwhile to scientifically look at this and define its value throughin vitro and in vivo assays. The applicant has developed the curiosityabout it and asked a number of questions: What possible importance cancamel urine have? Whether the component camel urine is having anyactivity on tumor formation by itself or some of its parts? Does thefresh urine have any effects on human cancer tissues? Could oneformulate this urine in a convenience form for human use? Does theformulation of urine enhance its activity or reduce it? Dose the camelurine contain microorganism? Is it safe to use it as a drug or does ithave toxicological effects on human diverse organs?

In addition, could one fractionate the lyophilized camel urine in orderto separate the bio-active fractions? To which part of the cell does thecamel urine react? What are the biochemical, biophysical, and biologicalevidences for its efficacy?

Camel's urine can be considered as an effective animal originsubstance/secretion with the capacity of improvement of some symptomsmainly associated with tumors formation in the body but it does needsubstantiation through scientific experimentation. Thus, the applicantconsidered it worthwhile to scientifically look at this and define itsvalues through in vitro and in vivo assays. The applicant in the firstinstance probed whether it contained anti-cancer agent since such aproperty would make it a highly useful natural substance. In relatedart, use of ‘piperine’ as a bioavailability enhancer has been describedin U.S. Pat. Nos. 5,616,593 and 5,972,382. Also cow's urine distillateor a dried fraction used for improving activity and bioavailability ofantibiotics drugs as described in later U.S. Pat. No. 6,896,907.

BRIEF SUMMARY OF THE INVENTION

Some of the above questions have been solved and addressed herein. Toanswer the first set of questions the applicant collected the camelurine (PM701) from natural pastures in various areas within Jeddah,Makah, Madinah and Riyadh governorate (Saudi Arabia) in different sizebottles at any time of the day. The urine was on CLED media and bloodagar in sterile condition and did not notice any growth ofmicroorganism. Tissue cultures of human cancer tissues and normaltissues were in studying the effect of (camel urine) PM701 on thebehavior of cancer cells and normal cells. PM701 appears to target thecancer cells and have anti-proliferative, apoptotic efficacy on them.

Surprisingly, the same PM701 exhibited nourishing effects on normalhealthy cells; this implies that PM701 have a selectively killing effecton cancer cells and reparative effect on normal dividing cells, theseresults leading to this invention. The novelty of the invention lies inthe fact revealed through precise experimentation that the PM701 actionand its effectiveness is achievable only in the range of concentrationwhich is literally in nano to micro-gram levels. That should be thereason for detection such a valuable potential of PM701 in targeted tothe cancer cells. The utilized of fresh PM701 remains non-acceptable andnon-convenience for human use therefore, the applicant also furtherlyophilized the liquid PM701 to obtain 0.2 g/ml of powder. were-examined the lyophilized PM701 on normal cells and diverse cancercells in both cell culture and animal models, which showed the sameanti-cancer efficacy and that was mediated by apoptosis as determined byan MTT test and electron microscopy examination.

The applicant thought of utilizing PM701 as an alternative drug forcancer therapy since it showed a target effect on cancer cells and noside effects on the normal tissues, but the amount of lyophilized PM701dosage per day was as a load in the body (46 capsules/day), ultimatelyleading to use one of the ways, which has been feasible for drasticallyreducing the daily dosage of this anti-cancer agent PM701 and increasingthe efficiency of the dosage activity too and has also high commercialimportance. Therefore the bio-guided fractionation approach was usedwith lyophilized PM701, which lead us to the isolation andidentification of the bio-active fractions, which is responsible for theanti-cancer efficacy observed with the whole urine.

For the purpose of the present invention the following terms are definedbelow.

The term “anti-cancer therapy” is intended to mean growthinhibition/eradication of primary tumors, stabilization of tumor growth,inhibition of metastasis formation, or prevention of tumor formation.Furthermore, anticancer activity also covers any combination between oursubstances and other known or investigational anticancer agents, inorder to improve the therapeutic efficacy of drugs.

The main purpose of this work is to reach an optimum alternative drugfor cancer treatment other than radiation or chemotherapy.

A mainstream approach is treating cancer with new methods other thanchemotherapy and radiotherapy, which have very bad side effects onnormal tissues.

Another objective of the invention is to provide new use of the PM701 asa selective anti-cancer agent.

In another objective of the invention is to provide a method forimproving activity of PM701 via its bio-active fractions.

Also another objective of the invention is to provide a process for theisolation of the active fractions form PM701 of camel urine or cowurine.

Still another objective of the invention is to provide the bio-activefractions of lyophilized PM701 as in an effective amount as a novelpharmaceutical composition (capsules).

The important objective of the invention is to prevent destruction ofnormal tissues during the process of cancer treatment.

To reach a protocol for treating cancer patients with availablesubstances and at a low cost.

The invention relates to a new use of known abundantly available camelurine and cow urine as anti-cancer agent and to provide the bio-activefractions of it that be useful in cancer treatment. In accordance withone aspect of the invention there is provided a novel anti-cancerpharmaceutical composition comprising an acceptable, effectiveanti-cancer amount of bio-active fractions of PM701. The inventedbio-active fractions targeted the cancer tissues without any sideeffects on the normal tissues.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

FIG. 1 is a chart of projected causes of death in Saudi Arabia.

FIG. 2 illustrates the chromatographic subfractionation of the organicsoluble fraction PMF led to the isolation of 7 subfractions referred toas G1-G7.

FIG. 3A illustrates a curve of the in vitro cytotoxic effect oflyophilized PM701 using human lung carcinoma cell line, A549 indifferent incubated periods comparing with non treated cancer cells.

FIG. 3B illustrates a curve of the in vitro non-cytotoxic effect oflyophilized PM701 using human foreskin cell line, HFS in differentincubated periods comparing with non treated normal cells.

FIG. 3C illustrates the in vitro cytotoxic effect of lyophilized PM701using human lung carcinoma cell line, A549.

FIG. 4A Illustrates a curve of the in vitro cytotoxic effect of PMFfraction using human lung carcinoma cell line, A549 in differentincubated periods comparing with non treated cancer cells.

FIG. 4B illustrates a curve of the in vitro non-cytotoxic effect of PMFfraction using human foreskin cell line, HFS in different incubatedperiods comparing with non treated normal cells.

FIG. 4C illustrates the in vitro cytotoxic effect of PMF fraction usinghuman lung carcinoma cell line, A549.

FIG. 5A illustrates a curve of the in vitro cytotoxic effect of PMFKsubfraction using human lung carcinoma cell line, A549 in differentincubated periods comparing with non treated cancer cells.

FIG. 5B illustrates a curve of the in vitro non-cytotoxic effect of PMFKsubfraction using human foreskin cell line, HFS in different incubatedperiods comparing with non treated normal cells.

FIG. 5C illustrates the in vitro cytotoxic effect of PMFK subfractionusing human lung carcinoma cell line, A549.

FIG. 6A illustrates the in vitro cytotoxic effect of −2 and −3concentration of PMF fraction using human lung carcinoma cell line,A549.

FIG. 6B illustrates the in vitro cytotoxic effect of −2 and −3concentration of PMFK subfraction using human lung carcinoma cell line,A549.

FIG. 7 illustrates ultrastructural morphological hallmarks features thatcharacterize apoptosis, as shown by chromatin condensation and membraneblebbing in treated cells with FIG. 8A illustrates an MTT test show theeffect of two different concentrations of PM701: −2 or high and −3 orlow on four different cell densities 1, 3, 6 and 10×103 cell/well.

FIG. 8B illustrates an MTT test shows the effect of PM701 on two typesof carcinogenic cells A549 and L1210 at 3×103/well.

FIG. 9 illustrates schematically the process of fractionation andsubfractionation.

DETAILED DESCRIPTION OF THE INVENTION

As shown in FIGS. 2-9, the present invention solves the problem ofsearching for and obtaining a plentifully, available, cheap, and naturalmaterial (PM701) as an anti-cancer agent with a higher and selectivepotency on cancer cells, obtained from camel urine or cow urine.Additionally, the bio-active fraction of PM701 was isolated, which iscoded PMF due to its highly selectively and highly cytotoxic propertieson cancer cells, which is responsible on the whole PM701 effect.Furthermore, PMF was subfractionated and led to purification of sevensubfractions, the seventh which is coded PMFK has cytotoxic propertieson cancer cells as much as PMF cytotoxic properties.

PMF and PMFK have cytotoxic and killing activities in vitro on lungcancer cell line A549 and leukemic cancer cell line L1210 withoutaffecting normal human foreskin cells HFS. More important was theobvious effects on inhibition the cancer cell division activity throughthe morphological hallmarks and biochemical features that characterizeapoptosis, as shown by loss of cell viability, chromatin condensation,and reducing metabolic activity using an MTT test.

In another embodiment, PM701 was used in vivo in treating animal modelsthat were inoculated with cancer cells; the result of in vivo testing isas satisfactory as in vitro effect at tissue culture level.

In an embodiment of the present invention a pharmaceutical compositioncomprising an effective amount of PMF or PMFK as bioactive anticancercompounds and pharmaceutically acceptable additives selected fromanticancer compounds.

In another embodiment, PMF and PMFK are used as bioavailabilityanticancer therapy directly or in combination with other anticancermolecules.

In yet another embodiment, PMF and PMFK are induced the apoptosis ofcancer cells.

In yet another embodiment, the PM701 fractionation helps us to isolatethe bio-active molecules, which act better on the target cancer cells bydecreasing its proliferation.

In yet another embodiment, the cancer cells may be lung, leukemia or anycancer cells.

In yet another embodiment, PMF used in vitro in the range between 80μg/ml to 800 μg/ml and PMFK used in the range between 0.2 μg/ml to 2μg/ml.

In still another embodiment, the bioactive fractions (PMF and PMFK)enhance the antiproliferative and apoptotic activities of anti-canceragents (PM701) for 2.8 folds.

The methodology followed by us for this screening included specificallydesigned in vitro and in vivo bioassays as described below. The celllines used in this invention were obtained from cell bank of TissueCulture Unit in King Fahd Medical Research Center (KFMRC).

Development of Powder Form

For enhancing the utility and convenience of application of PM701,liquid PM701 was lyophilized to reach a solid form. The applicantfurther fractionated the solid form to obtain the bioactive fraction(s),which is also free of the typical smell of camel urine that it is morereadily acceptable to the humans. For this purpose the lyophilized PM701was fractionated as described by the following procedure:

Step 1: PM701 was collected in the stainless steel container directlyfrom the camel, which is maintained in hygienic environment.

Step 2: 90 gm of liquid PM701 was added to Microcrystalline cellulose(10 gm). This will give 100 gm of a mixture, that frozen at −80° C. inPyrex apparatus for 20-24 hrs.

Steps 3: 100 gm of the mixture is lyophilized in the lyophilizer at roomtemperature for 5 days to obtain 20 gm of solid form PM701 .

Step 4: Mixtures leave in a descator with calcium chloride, in presentof vacuum pressure for one day at room temperature

Step 5: The lyophilized PM701 is packed in fridge in a sterilized glasscontainer for further use.

Fractionation of a Lyophilized PM701 Separation of Active Ingredients

The following steps were performed.

i. Solvent Extraction Method for Fractionation

Step 1: A sample of 5 mg of lyophilized PM701 was sonicated withmethanol three times each 30 ml to give about 750 mg of methanolfraction, which is called (PMF).

ii. Molecular Sieving Method for Subfractionation

Step 1: Subfractionation of PMF using column of different type of gelwith different sieving capacity (e.g. Sephadex LH-20, Sephadex-25, -50,. . . ).

Step 2: A sample of 1.5 mg of The methanol fraction (PMF) waschromatographed on Silica gel column and was eluted with followingsolvents systems each 250 ml, chloroform, 10% methanol in chloroform,20% methanol in chloroform, 30% methanol in chloroform, 40% methanol inchloroform, 60% methanol in chloroform followed with methanol.

Step 3: The seven subfractions are purified and individual sub fractionsare separated by high-performance liquid chromatography.

Step 4: All subfractions, which comes out of column were tested forsimilar activity as that of PM701 at the tissue culture level.

The organic solvent soluble fractions and subfractions were found tohave a strong antiproliferative activity in a panel of human cancer celllines derived from lung and leukemia. In vitro, the fractions andsubfractions demonstrate antiproliferative and antiapoptotic activitiesin tissue culture experiments, as shown in FIGS. 2 and 9.

Assay for In Vitro Dose Determination

a. The optimum inhibitory concentration of PM701 is evaluated anddetermined against different cancer cells, lung cancer cell line (A549),leukemic cell line (L1012) through the in vitro tissue cultureexperiments.

b. The optimum inhibitory concentration of PM701 showing a cytotoxiceffect on cancer cells while there is no any cytotoxic effect on normalcells, suggesting that PM701 can act selectively on cancer cells whilenourishing normal cells.

1 ml dissolved PM701 or PMF or PMFK in 10 ml standard media, which iscalled −1 (high).

1 ml dissolved PM701 or PMF or PMFK in 100 ml standard media, which iscalled −2

1 ml dissolved PM701 or PMF or PMFK in 1,000 ml standard media, which iscalled −3 (mid).

1 ml dissolved PM701 or PMF or PMFK in 10,000 ml standard media, whichis called −4.

1 ml dissolved PM701 or PMF or PMFK in 100,000 ml standard media, whichis called −5 (low).

The in vitro experiments showed that the best effect observed when used−2 and −3 concentrations, so we fixed the in vitro and in vivo dosesusing these medium concentration.

In Vitro Antiproliferative Activity Assay (Cell Culture)

1. Cell Lines and Cell Culture

a. A549 (Lung cancer commercial cell line obtained from King FahdMedical Research Center (KFMRC) is inoculated at a density of about0.5×105 cells in MEM medium in the wells of 24 well plate. L1210 cellswere prepared by the same way in RPMI (1640) supplemented with 10%heat-inactivated fetal calf serum.

b. This is replaced with fresh medium after 24 hours in each well.

c. The test component(s) is added at desired concentrations in differentwells just after the medium replacement.

d. Observations are recorded on the cell count after 0, 24, 48, and 72hours for which the following steps are required.

i. The medium is removed from the wells.

ii. The wells are rinsed with 1 ml PBS (Phosphate buffer saline).

iii. To each well 500 μl of freshly prepared trypsin (0.1% in PBS)solution is added.

iv. Typsin solution is removed after 30 seconds and the plate is gentlytapped till the cells are released from the plate surface.

v. Fresh 1 ml of growth medium is added and agitated with a pipette toobtain a cell suspension.

vi. Cell suspension was prepared (1:1): 20 ml of cells with 20 ml of(0.4%) Trypan blue, 10 μl of cell suspension is taken on thehaemocytometer and a cover glass is placed over the counting chamber.

vii. The number of viable cells is counted in 5 big squares and thereadings are taken from 5 microscopic fields to determine the average.

viii. The cell count (titer per ml) in the original sample is thencalculated as average count×104.

Composition of Minimum Essential Medium (MEM)

MEM powder (ICN)=9.95 g, NaHCO3 (Powder)=2.2 g, glutamine (Powder)=0.3 gL, Non essential amino acid (100×)=10 ml, Hepes (100×) solution=10 ml,antibiotic mix (Penicillin+Streptomycin)=10 ml, deionzed-distilledwater=1 liter.

Stirrer for 1 hrs at room temperature, PH (6.8-7.4).

Sterile-filtered through a 0.22 μm filter and stored at +4 co.

FIG. 3A illustrates a curve of the in vitro cytotoxic effect oflyophilized PM701 using human lung carcinoma cell line, A549 indifferent incubated periods comparing with non treated cancer cells.

FIG. 3B illustrates a curve of the in vitro non-cytotoxic effect oflyophilized PM701 using vero cell line in different incubated periodscomparing with non treated normal cells.

FIG. 3C illustrates the effect of lyophilized PM701 on the cellmorphology of human lung carcinoma cell line, A549. Cancer cells A549imaged (40×) after incubation for 24 h, fixed and stained with Coomassieblue (a) in PM 701. Note the damage of cells as compared with thecontrol cells that were incubated in MEM media (b).

FIG. 4A Illustrates a curve of the in vitro cytotoxic effect of PMFfraction using human lung carcinoma cell line, A549 in differentincubated periods comparing with non treated cancer cells.

FIG. 4B illustrates a curve of the in vitro non-cytotoxic effect of PMFfraction using human foreskin cell line, HFS in different incubatedperiods comparing with non treated normal cells.

FIG. 4C Illustrates the effect of PMF fraction on the cell morphology ofhuman lung carcinoma cell line, A549, a. treated cells; b. non treatedcells (40×).

FIG. 5A illustrates a curve of the in vitro cytotoxic effect of PMFKsubfraction using human lung carcinoma cell line, A549 in differentincubated periods comparing with non treated cancer cells.

FIG. 5B illustrates a curve of the in vitro non-cytotoxic effect of PMFKsubfraction using human foreskin cell line, HFS in different incubatedperiods comparing with non treated normal cells.

FIG. 5C illustrates the effect of PMFK subfraction on the cellmorphology of human lung carcinoma cell line, A549, a. treated cells; b.non treated cells (20×).

FIG. 6 illustrates ultrastructural morphological hallmarks features thatcharacterize apoptosis, as shown by chromatin condensation and membraneblebbing in treated cells with PM701.

Cytotoxicity Assay

The MTT test is used for determination of the cytotoxicity or theanticarcinogenic effects of PM701 on two types of cancer cells, A549 andL1210. This test measures the cell viability as a percentage of controluntreated cells.

An MTT assay was performed to evaluate the growth effects of PM 701. TheMTT assay is calorimetric assay based on the tetrazolium salt MTT thatdetects cell viability. Dissolved MTT is converted to an insolublepurple formazan by cleavage of the tetrazolium ring by dehydrogenaseenzymes in living but not dead cells.

MTT was dissolved in phosphate buffered saline (PBS) at 5 mg/ml andfiltered through a 0.22 μm filter to sterilize and remove the smallamount of insoluble residue then stored at 2-8 CO for frequent use.Stock solution of MTT is added to each culture being assayed to equalone tenth the original culture volume.

Using isopropanol is measured by spectrophotometrically yieldingabsorbance as a function of concentration of converted dye.

Exponentially growing cells (3×103 cells/100 μl) were seeded in 96-wellplates and incubated for 24 h. Cells were then treated continuously withthe various fractions and subtractions. At a selected time, 10 μl ofstock MTT solution was added to all wells for the assay. After a furtherperiod of incubation (4 hours), the medium was aspirated from the wellsas completely as possible without disturbing the formazan crystals.Then, 100 μl of isopropanol is added to each well for dissolving theresulting precipitate. The concentration of the dye is then measured at570 nm on plate reader (Microplate Reader Model 450; Bio-Rad). Theoptical density obtained is directly related to the viability of cells.

The MTT assay distinguishes between viable and non-viable cells on thebasis that physiologically active mitochondria metabolizes the MTT onlyin viable cells. The IC50 was calculated as the concentration of drugcausing a 50% inhibition in the absorbance compared to cells treatedwith solvent alone.

FIG. 8A illustrates an MTT test show the effect of two differentconcentrations of PM701: −2 or high and −3 or low on four different celldensities 1, 3, 6 and 10×103 cell/well.

FIG. 8B illustrates an MTT test shows the effect of PM701 on two typesof carcinogenic cells A549 and L1210 at 3×103/well.

Results

Methanol soluble fraction (PMF) but not water soluble fractions, wasfound to have a potent antiproliferative activity in A549 and L1210 celllines. Further chromatographic subfractionation of these organic solubleextract led to the isolation of 7 fractions referred to as G1-G7, asshown in FIG. 2 which illustrates the chromatographic subfractionationof the organic soluble fraction PMF led to the isolation of 7subfractions referred to as G1-G7. Solvent system: CHCl3-MeOH-Water65:35:6. Spray Reagent: P-anisaldehyde Reagent. Heating at 110° C. for 5min.

The morphological changes of treated cells with lyophilized PM701characterize apoptosis, as shown by loss of cell viability, membraneblebbing, and chromatin condensation. and shown in FIGS. 7A and 7B. FIG.7A illustrates the in vitro cytotoxic effect of −2 and −3 concentrationof PM701 using mice leukemia cells, L1210. FIG. 7B illustrates theeffect of PM 701 on the cell morphology of mice leukemia cells, L1210,a. treated cells; b. non treated cells. Note the normal cell size(arrows) (40×).

Methanol soluble fraction PMF was found to have a good anticanceractivity carcinoma cell lines. A dose relationship was also observed, asshown in FIG. 6.

Alternative Embodiments

Pharmaceutical compositions having anticancer properties can also beobtained from cow urine in the same manner described herein forobtaining such pharmaceutical compositions from camel urine. In thealternative embodiment, the pharmaceutical composition includes a leastone anticancer agent; and a cow urine distillate or a dried fraction(GM-IV) obtained from cow urine distillate. The cow urine distillate maybe present in a concentration range of 0.001 pl/ml to 100 pl/ml. Thepharmaceutical composition may include Taxol. The dried fraction (GM-IV)of the cow urine distillate may be obtained by lyophilization, and mayhave the following physical characteristics: a white color, a solidcrystalline form, water solubility, a melting point above 400° C., aspecific gravity of 1.006 and an RF value in methanol: chloroform(50:50) phase 0.65. Preferably, the dried fraction (GM-IV) obtained fromthe cow urine distillate is devoid of a cow urine smell.

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprinciples of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention pertains and as may be applied to theessential features hereinbefore set forth, and as follows in the scopeof the appended claims.

Still more interesting observation is that the in vivo study on theanimal models, which indicated that (PM 701) has the ability to limitcancer progression in treated animals by at least 3 folds, which meansthat it has a favorable antimitotic effect. Further patent in thisdirection have been in progress.

1. A novel use of camel urine coded as PM701 as an anti-cancer agent. 2.A pharmaceutical composition comprising: at least one anticancer agent,and a camel urine lyophilized or a dried fractions (PMF and PMFK)obtained from camel urine lyophilized.
 3. The composition of claim 2,wherein said camel urine is the adult single-humped Arabian camelCamelus dromedarius.
 4. The composition of claim 2, wherein thecomposition is obtained in a powder form by the lyophilization of PM701in the range of 20 g for each 100 ml of fresh liquid PM701.
 5. Thecomposition of claim 4, comprising the following physicalcharacteristics: a yellowish color, a solid crystalline form, a sharp(offensive) odor, a pH is 8.3, water insoluble and a specific gravity of1.045.
 6. The composition of claim 4 wherein the composition exhibits atarget selective anti-cancer effect in causing antiproliferative andapoptotic activities in vitro on diverse cancer tissues representing twodifferent types of cancers (human lung cells (A549) and mice leukemiccells (L1210)) and nourishing effects on human normal tissues of humanforeskin and vero cells.
 7. The composition of claim 4, wherein thecomposition is fractionated by a number of trials in order to isolatethe most effective bio-active fraction (PMF) through the bio-guidedfractionation.
 8. The composition of claim 7 obtained by fractionationin the range 150 mg/g of lyophilized PM701 .
 9. The composition of claim7, wherein the fraction isolated from the lyophilized PM701 is subjectedfor further subfractionation in solvent systems each 250 ml consistingessentially of chloroform and methanol to obtain seven subfractions,with a most effective subfraction being called PMFK.
 10. The compositionof claim 9 obtained by subfractionation in the range 108.7 mg/g of PMF.11. The composition of claim 10 comprising about 108.7 mg/g of thecomposition including about 150 mg/g of lyophilized PM701.
 12. Thecomposition of claim 7, wherein concentration effects on the cellviability and proliferation of the cancer cells was at concentrations inthe range of 80 μg and 800 μg.
 13. The composition of claim 9, whereinconcentration effects on the cell viability and proliferation of thecancer cells was at concentrations in the range of 0.2 μg and 2 μg. 14.The composition of claim 7, comprising an effective amount of thecomposition in the amount of 435 mg/capsule with 7 capsules/day.
 15. Thecomposition of claim 9, comprising an effective amount of thecomposition in the amount of 435 mg/capsule with 5 capsules/day.
 16. Thecomposition of claim 2, wherein the dried PM 701 of the camel urinelyophilized can be obtained by distillation.
 17. The composition ofclaim 7, wherein the dried fractions (PMF and PMFK) obtained from thecamel urine lyophilized are devoid of a camel urine smell.
 18. Thecomposition of claim 9, wherein the dried fractions (PMF and PMFK)obtained from the camel urine lyophilized are devoid of a camel urinesmell.
 19. A pharmaceutical composition comprising: a least oneanticancer agent; and a cow urine distillate or a dried fraction (GM-IV)obtained from cow urine distillate.
 20. The pharmaceutical compositionof claim 19, wherein the cow urine distillate is present in aconcentration range of 0.001 pl/ml to 100 pl/ml.
 21. The pharmaceuticalcomposition of claim 19, further comprising Taxol.
 22. Thepharmaceutical composition of claim 19, wherein the dried fraction(GM-IV) of the cow urine distillate is obtained by lyophilization. 23.The composition of claim 22, wherein the dried fraction (GM-IV) obtainedfrom the cow urine distillate has the following physicalcharacteristics: a white color, a solid crystalline form, watersolubility, a melting point above 400° C., a specific gravity of 1.006and an RF value in methanol: chloroform (50:50) phase 0.65.
 24. Thecomposition of claim 22, wherein the dried fraction (GM-IV) obtainedfrom the cow urine distillate is devoid of a cow urine smell.